Development Of A Novel Cell Culture Medium For AAV Production And Process Optimization

Adeno-associated virus (AAV) has become a prominent vector to treat life-threatening genetic diseases. As AAV therapeutic demand increases, so does the need for large quantities of high-titer AAV. A versatile GMP medium is vital to achieving the high AAV titers essential to support this rapidly growing industry. In addition, the Gene Therapy industry relies on various commercially available HEK293 cell lines for AAV production, and finding the optimal medium is critical to achieving cell growth and AAV titer goals.

Here, we describe the development of the Cellvento® 4HEK Medium, a novel cell culture medium for AAV production with multiple HEK293 lineages. To maximize AAV titers across multiple HEK293 cell lines, the medium development process was performed with three different HEK293 cell lines, each one with diverse growth rates. Subsequently, the medium prototype was optimized to provide the highest AAV titer across the different cell lines. Following media development, MilliporeSigma sought to optimize the PEI transfection and bioreactor parameters. With this approach, the highest VCD and AAV titers can be achieved without the bottleneck of a non-optimized step in the AAV production process.